title: Fetal mesenchymal stromal cells from cryopreserved human chorionic villi: cytogenetic and molecular analysis of genome stability in long-term cultures.
abstract: OBJECTIVE
  First-trimester chorionic villi (CV) are an attractive source of human mesenchymal stromal cells (hMSC) for possible applications in cellular therapy and regenerative medicine. Human MSC from CV were monitored for genetic stability in long-term cultures.
  
  
  METHODS
  We set up a good manufacturing practice cryopreservation procedure for small amounts of native CV samples. After isolation, hMSC were in vitro cultured and analyzed for biological end points. Genome stability at different passages of expansion was explored by karyotype, genome-wide array-comparative genomic hybridization and microsatellite genotyping.
  
  
  RESULTS
  Growth curve analysis revealed a high proliferative potential of CV-derived cells. Immunophenotyping showed expression of typical MSC markers and absence of hematopoietic markers. Analysis of multilineage potential demonstrated efficient differentiation into adipocytes, osteocytes, chondrocytes and induction of neuro-glial commitment. In angiogenic experiments, differentiation in endothelial cells was detected by in vitro Matrigel assay after vascular endothelial growth factor stimulation. Data obtained from karyotyping, array-comparative genomic hybridization and microsatellite genotyping comparing early with late DNA passages did not show any genomic variation at least up to passage 10. Aneuploid clones appeared in four of 14 cases at latest passages, immediately before culture growth arrest.
  
  
  CONCLUSIONS
  Our findings indicate that hCV-MSC are genetically stable in long-term cultures at least up to passage 10 and that it is possible to achieve clinically relevant amounts of hCV-MSC even after few stages of expansion. Genome abnormalities at higher passages can occasionally occur and are always associated with spontaneous growth arrest. Under these circumstances, hCV-MSC could be suitable for therapeutic purposes.
authors.affiliation: TOMA Advanced Biomedical Assays S.p.A., Laboratorio di Ricerca e Sviluppo, Busto Arsizio, Italy. Electronic address: manurosel@libero.it.
authors.forename: Emanuela Anna
authors.initials: EA
authors.lastname: Roselli
authors.forename: Silvia
authors.initials: S
authors.lastname: Lazzati
authors.forename: Federico
authors.initials: F
authors.lastname: Iseppon
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Manganini
authors.forename: Livia
authors.initials: L
authors.lastname: Marcato
authors.forename: Marzia Bruna
authors.initials: MB
authors.lastname: Gariboldi
authors.forename: Federico
authors.initials: F
authors.lastname: Maggi
authors.forename: Francesca Romana
authors.initials: FR
authors.lastname: Grati
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Simoni
mesh_terms: D017667:Adipocytes / Q000166:cytology; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D064987:Cell- and Tissue-Based Therapy; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology; D002824:Chorionic Villi / Q000196:embryology*; D055028:Comparative Genomic Hybridization; D015925:Cryopreservation; D005260:Female; D014644:Genetic Variation; D042822:Genomic Instability / Q000235:genetics*; D006801:Humans; D016130:Immunophenotyping; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D018895:Microsatellite Repeats / Q000235:genetics; D010011:Osteocytes / Q000166:cytology; D011247:Pregnancy; D011261:Pregnancy Trimester, First; D023822:Tissue Engineering
keywords: array-comparative genomic hybridization; chorionic villi; genomic instability; human placenta; mesenchymal stromal cells